High-Throughput Screening of T Cell Cytotoxic Events by Biomass Profiling

Abstract

Adoptive immunotherapies against cancer, in which cytotoxic, CD8+ T cells engineered to express T cell receptors (TCRs) targeting cancer-associated antigens are transplanted into a patient, have shown dramatic promise in clinical trials. A major impediment to the widespread use of this technique for treatment of diverse cancers is the lack of a fast approach for the identification of TCRs from patient samples. In this talk, we present a method for high-throughput screening of T cell/target cell interactions by measurements of cell biomass. This live cell approach is label-free and allows cells to be recovered for downstream analysis. To ensure specificity, three parameters are tracked: target cell appearance, target cell mass loss during cell death, and T cell mass during and after the cytotoxic event (Figure panels a-c). Our results demonstrate, for the first time, the kinetics of T cell mass increase during activation. Finally, we will present an extension of this method to a microfabricated microwell format for the screening of patient samples and discovery of novel TCRs.View Large Image | View Hi-Res Image | Download PowerPoint Slide